Compositions for the reversal of stratum granulosum in keratinization disorders

ABSTRACT

Compositions that, when used topically in a therapeutically effective amount, are safe and effective for the regression of stratum granulosum in patients exhibiting keratinization disorders, such as psoriasis. The compositions, which can be formulated as ointments, oils, soaps and shampoos, comprise a non-aqueous extract of Wrightia tinctoria, an extract of Cocos nucifera, and pharmaceutically or cosmetically acceptable excipients suitable for topical use.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application, filed under 35 U.S.C. 120, is acontinuation-in-part of U.S. non-provisional patent application Ser. No.11/731,345 entitled “Compositions for the Reversal of Stratum Granulosumin Keratinization Disorders” filed on Mar. 31, 2007, and as such claimspriority benefit under 35 U.S.C. 120 of this application. The presentapplication is related to U.S. non-provisional applications Ser. No.11/288,923 filed on Nov. 28, 2005; Ser. No. 12/009,799 filed on Jul. 3,2008; and Ser. No. 12/286,564 files on Sep. 30, 2008. The disclosures ofthese applications are incorporated herein by reference in theirentireties.

STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

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THE NAMES OF THE PARTIES OF A JOINT RESEARCH AGREEMENT

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INCORPORATION-BY-REFERENCE OF MATERIAL SUBMITTED ON A COMPACT DISC

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FIELD OF THE INVENTION

The present invention relates to compositions for the reversal ofstratum granulosum in keratinization disorders. In particular, thepresent invention relates to herbal compositions for the regression andreversal of stratum granulosum in keratinization disorders in psoriaticlesions.

BRIEF DESCRIPTION OF THE BACKGROUND ART

The skin is made up of two primary layers that differ in function,thickness, and strength. From outside to inside, they are the epidermisand its sublayers, and the dermis, after which is found subcutaneoustissue, or the hypodermis.

The epidermis, the outermost layer of skin, is thin but complex.Melanin, which is responsible for skin pigmentation, is found throughoutthe epidermis. The epidermis also keratinizes to produce nails, hair,sweat, and to regenerate.

Keratinization, the maturation and migration of skin cells, begins inthe innermost layer of the epidermis, the stratum germinativum [see itemE in FIG. 1]. These cells, called keratinocytes, accumulate and moveoutward toward the next epidermis layer, the stratum spinosum [see itemD in FIG. 1], where they become dense. The next layer, known as thestratum granulosum [see item C in FIG. 1] layer, contains 1 to 3 rows offlattened cells whose cytoplasm contain small granules. The granulescontain proteins being transformed into the waterproofing proteinkeratin. It is in this layer that one finds glycolipids and a thickeningof the membrane. A protein called filigrin is made in this layer and isput in the granules. In this layer, cells lose their nuclei. In thecytoplasm, there are keratohyalin granules as well as membrane-coatinggranules which expel their lipid contents into the intercellular spaces.Lipids assist in the formation of water barriers among the cells of theskin, which, in turn, help to ensure body moisturization. At this point,the cell also becomes flattened, or horny, and the nucleus disappears;what remains is keratin. In the next layer, the stratum lucidum [seeitem B in FIG. 1], the cell is prepared to move into its final sublayerwith the addition of melanin granules. Then, sudden changes in enzymefunction cause cell death. The products of this ongoing process form thestratum corneum [see item A in FIG. 1], which is the outermost epidurallayer consisting of neatly packed dead horny cells.

Keratinization disorders in the stratum granulosum layer in theepidermis can often lead to clinically significant skin manifestations.One common disorder includes thinning of the stratum granulosum layerdue to malfunctioning of the keratinization process leading to reductionin the moisture barrier properties of the stratum granulosum layer. Inaddition, for example, over activated keratinocytes actively producingand secreting pro-angiogenic factors in the form of growth factors orcytokines can result in increased blood vessel formation in thepapillary dermis which may sometime extend into the epidermis. Epidermalmicrovascular proliferation ultimately leads to epidermal keratinocytehyperproliferation, thickening of the epidermis with parakeratosis ofthe stratum corneum and inflammatory infiltrate around the blood vesselsin the papillary dermis [see FIG. 1]. The microvascular changes are alsocharacterized by increased tortuosity of dermal capillary loops whichprecedes the development of epidermal hyperplasia. Mitotic activity inthe basal and suprabasal cells are greatly increased [Dr. George Jacob,Seminar on Psoriasis, Dubai, January 2001]. Cellular invasion takesplace, particularly in the suprapapillary region to form the Munro‘micro abscess’ which are extruded in the horny layer or they maycollect in disintegrated malphigian cells, the cytoplasm of which hadbeen lysed to form the multilocular or stratum granulosum of Kogoj.

Stratum granulosums of Kogoj are multilocular pustules in the upperstratum malpighii within a sponge-like network made up of flattenedkeratinocytes [M. S. Stone and T. L. Ray, DermPath Tutor, Department ofDermatology, Iowa College of Medicine, September 1995]. They are seen inpsoriasis, Reiter's disease, geographic tongue and rarely incandidiasis. Histological studies, including immunocytochemistry,routine histology and electron microscopy have clearly established thatalterations in the blood vessel formation of the skin discussed aboveare a prominent feature of psoriasis and there is a marked increase incutaneous blood active edge of the psoriatic plaque [Braverman I M, YenA. Ultrastructure of the capillary loops in the dermal papillae ofpsoriasis. J Invest Dermatol 1977: 68: 53-60].

Numerous therapies in the field of allopathy medicine have been reportedin reducing a stratum granulosum disorder, especially in relation topsoriasis:

-   -   Treatment of psoriasis—Part 1—Topical Therapy and Phototherapy,        Mark Lebwohl, MD, et al, American Academy of Dermatology,        October 2001 Vol 45 (4).    -   Treatment of psoriasis—Part 2—Systemic Therapies, Mark Lebwohl,        MD, et al, American Academy of Dermatology, November 2001 Vol        45(5).    -   The immunological basis for the treatment of psoriasis with new        biological agents. James. G. Krueger, M.D, American Academy of        Dermatology, June 2002 Vol 46(1) Pages 1-26.    -   New psoriasis treatments based upon a deeper understanding of        the pathogenesis of psoriasis vulgaris and psoriatic arthritis.        Jeffrey P. Callen et al, American Academy of Dermatology, August        2003 Vol 49(5) Pages 351-356.

However, most of these therapies provide only temporary symptomaticrelief and are either unsatisfactory or very expensive and areassociated with either short term or long term undesired side effectprofiles. [National Psoriasis Conference, Boston Plaza Hotel, Aug. 5-8,2005, Boston, Mass., USA.] In addition, it is possible for thetreatments to lessen the severity of one or more of the manifestationsof psoriasis, but not all. These manifestations include but are notlimited to stratum granulosum of Kogoj, dermal vessel tortuosity, andspongiform pustules.

It is well known that herbal formulations often have fewer undesirableside effect profiles and hence provide a viable alternative therapy tomanage the keratinization disorders that exhibit stratum granulosum.Research efforts to develop herbal formulations to treat theseconditions have been on the rise and there is a continuing need todevelop herbal formulations to treat stratum granulosum inkeratinization disorders with minimal or no side effects:

-   -   Chopra, R. N., Nayar, S. C., and Chopra I. C., Glossary of        Indian Medicinal Plants, C.S.I.R., P.259, 1956.    -   Murugesa Mudaliar, K. S., Gunapadam (Material Medica) Vegetable        Section, Govt. of TamilNadu, P. 527 (1969).    -   Venkatarajan, S., Sarabendra Vaithiya Muraigal, P. 160, 161 &        167 (1965).    -   Wealth of India, Raw Materials, Vol. X, P. 588-590, CSIR., New        Delhi (1976).    -   Yugimuni Vaidya Chintamani (800) Stanza 494-518, B. Rathina        Nayakar & Sons, Madras, India.    -   Nair, C. P. R., Kurup, P. B., Pillai, K. G. B., Geetha, A., and        Ramiah, N., Effect of Nimbidin in Psoriasis, Indian Medical        Journal, October 1978.

The invention provides herbal formulations, developed by adermatologist, based on Wrightia tinctoria and an extract of Cocosnucifera which, when used as directed in therapeutically effectiveamounts, have been clinically proven to be safe and efficacious inreversing or regressing stratum granulosum in keratinization disordersin humans in need of treatment. The definition of the term herb, as itis used here is taken from the definition provided by the Herb Societyof America: The tem herb refers to a wide range of plants, includingperennials, trees, shrubs, annuals, vines, and more primitive plants,such as ferns, mosses, algae, lichens, and fungi. The herbs are valuedfor their flavor, fragrance, medicinal and healthful qualities, economicand industrial uses, pesticidal properties, and coloring materials(dyes).” [Bown, Deni. The Herb Society of America New encyclopedia ofHerbs and Their Uses. New York: Dorling Kindersley, 2001, p. 18]

SUMMARY OF THE INVENTION

It is an object of the present invention to provide a composition that,when used topically as directed, is suitable for the reversal of stratumgranulosum in keratinization disorders. The composition can beformulated, for example, as an ointment, an oil, a soap or a shampoo.The composition is comprised of efficacious amounts of a non-aqueousextract of Wrightia tinctoria, an extract of Cocos nucifera and suitablepharmaceutically or cosmetically acceptable excipients designated fortopical use in humans.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1: Illustration of the stratum granulosum layer in the epidermis

FIG. 2: Micrographs illustrating stratum granulosum in keratinizationdisorders—before and after treatment

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to formulations, which unexpectedlyprovide statistically superior efficacy to allopathy controlformulations in reversal of stratum granulosum disorder when used intherapeutically effective amounts. They are proven safe to use in humanswhen used topically as directed.

It is an object of the invention to provide a composition that comprisesa non-aqueous extract of Wrightia tinctoria, an extract of Cocosnucifera, and pharmaceutically or cosmetically acceptable excipientssuitable for topical use. The composition can be formulated as anointment, an oil, a soap and a shampoo, and, when a therapeutic amountis applied topically to the affected area, in the treatment of stratumgranulosum in keratinization disorders and is safe in humans.

The herbal extract in the topical composition for the regression ofstratum granulosum in keratinization disorders is derived from theWrightia tinctoria plant, especially the leaves, twigs, flowers, fruitsor a combination of these parts of the plant. Wrightia tinctoria is anapocynaceae tree growing throughout India. Its flowers are white andfragrant.

It is another object of the invention to provide a process for preparingthe non-aqueous extract of Wrightia tinctoria. The non-aqueous extractis prepared at ambient temperature by cleaning and pulverizing theselected parts of the Wrightia tinctoria plant and soaking them in anon-aqueous oil medium containing coconut oil. Care should be taken toadd sufficient oil medium to ensure that the plant material iscompletely submerged. The plant material/oil medium is then irradiatedwith a light source in the spectrum range of 300-1100 nm for a period ofapproximately 5 days. During this time the herbal ingredients areallowed to extract into the non-aqueous oil medium. At the end of theextraction, the oil medium is a purplish brown color. It is thenfiltered and the filtrate is stored for further processing as thenon-aqueous herbal extract of Wrightia tinctoria.

Other herbal extracts may optionally be included in the formulation,including Melia azadirachta Linn oil (Neem oil), which has beendocumented to have beneficial skin effects [Nair et al., 1978]. Thetopical composition of the invention for the reversal of stratumgranulosum in keratinization disorders optionally comprises an extractof the active herbal ingredient mentioned above in the extraction mediumin the amount from 1% to 20% by weight.

The herbal extract of Cocos nucifera in the composition of thisinvention is derived from the copra of the coconut. The copra of thecoconut is dried and then processed by grinding and pressing to extractthe oil, which is then purified and stabilized. The composition of thepresent invention comprises the herbal extract (the oil) of Cocosnucifera present in the amount of 40% to 80% by weight.

It is an object of the invention to provide formulations for topical useby further compounding the compositions of the invention withingredients mentioned herein to prepare formulations, including but notlimited to, an ointment, an oil, a liquid soap and a shampoo.

The ointment formulation of the herbal composition of the inventionsuitable, when used topically in a therapeutically effective amount, forthe reversal of stratum granulosum in keratinization disorders, includespharmaceutically acceptable excipients such as beeswax, paraffin(liquid, soft and hard), and other standard ointment bases or theirequivalents to optimize the use characteristics of the formulations,such as consistency and spreadability, as well as manufacturability andstability. The ointment composition comprises one or more of theexcipients including beeswax, optimally present in the amount of 1 to 5%by weight; paraffin, optimally present in the amount of 5 to 40% byweight; and standard ointment bases, optimally present in the amount of5 to 50% by weight.

The oil formulation of the herbal composition of the invention suitable,when used topically in a therapeutically effective amount, for thereversal of stratum granulosum in keratinization disorders, includespharmaceutically acceptable excipients such as vegetable oil, animaloil, and synthetic oils such as mineral oil and liquid paraffin or theirequivalents to optimize the use characteristics of the formulations,such as consistency and spreadability, as well as manufacturability andstability. Preferentially, the oil composition comprises excipients,such as coconut oil, present in the amount of 70 to 95% by weight.

The liquid soap formulation of the herbal composition of the inventionsuitable, when used topically in a therapeutically effective amount, forthe reversal of stratum granulosum in keratinization disorders, includespharmaceutically acceptable excipients, including but not limited to:water, surface active agents, thickeners and viscosity enhancers, foamboosters, and stabilizers to optimize the use characteristics, such asconsistency, cleaning, spreadability and foaming, as well asmanufacturability and stability. The liquid soap formulation of thepresent invention preferentially comprises excipients such as waterpresent in the amount of 60 to 85% by weight; surface active agentspresent in the amount of 5 to 40% by weight; thickeners or viscosityenhancers present in the amount of 0.5 to 8% by weight; foam boosterspresent in the amount of 1 to 4% by weight; and stabilizers present inthe amount of 0.5 to 2% by weight.

The shampoo formulation of the herbal composition of the inventionsuitable, when used topically in a therapeutically effective amount, forthe reversal of stratum granulosum in keratinization disorders, includespharmaceutically acceptable excipients, comprising water, surface activeagents, thickeners or viscosity enhancers, foam boosters, andstabilizers to optimize the use characteristics such as consistency,cleaning, spreadability and foaming, as well as manufacturability andstability. The shampoo composition of the present inventionpreferentially comprises excipients including water present in theamount of 50 to 85% by weight; surface active agents present in theamount of 10 to 30% by weight; thickeners or viscosity enhancers presentin the amount of 2 to 8% by weight; foam boosters present in the amountof 2 to 6% by weight; and stabilizers present in the amount of 0.5 to 2%by weight.

In addition, the ointment, oil, liquid soap and shampoo formulations ofthe herbal composition of the invention suitable, when used topically ina therapeutically effective amount, for the reversal of stratumgranulosum in keratinization disorders, optionally comprisepreservatives, coloring agents and fragrances as needed, wherein thepreservatives, coloring agents and fragrances are present in the amountof 0 to 5 total weight %.

Safety and efficacy studies were conducted on subjects exhibiting ofstratum granulosum in keratinization disorders using topicalformulations of the herbal composition of the present inventiondescribed above, containing a non-aqueous herbal extract of Wrightiatinctoria, an herbal extract of Cocos nucifera and pharmaceutically orcosmetically acceptable excipients. Patients suffering from chronicinflammatory skin conditions selected for the study exhibited stratumgranulosum in keratinization disorders in the form of psoriatic lesions.Psoriasis is a representative example of a condition exhibiting stratumgranulosum in keratinization disorders. The results are illustrated bythe following example.

EXAMPLE

Twenty patients were enrolled in a clinical study and were divided intotwo groups of 10 patients each. Group I was treated with the herbalformulation (see Table 1 for details) once daily and Group II wastreated with an allopathy control formulation (see Table 2 for details)once daily. All patients recruited were screened and were determined tobe suffering from stratum granulosum in keratinization disorders. Allpatients were psoriasis patients.

TABLE 1 Herbal Ointment Formula of Invention No. Ingredient Quantity 1Wrightia Tinctoria 5% 2 Cocos Nucifera 65%  3 Bees Wax 6% 4 LiquidParaffin 24% 

TABLE 2 Dithranol Ointment (Allopathy Control) No. Ingredient Quantity 1Dithranol 1% 2 Standard Ointment Base QS

Assignment of patients to treatment groups was randomized as perstandard statistical methods to minimize bias in the study. Patientswere enrolled in the study on a first come, first served basis andassigned a subject number sequentially. The assignment of each patientto the treatment group was determined by the randomization list providedby the statistician.

Each patient voluntarily enrolled in the study and received thetreatment for 8 weeks. Skin biopsies at the treatment site were takenfrom all patients at the beginning (T0) and end of the study (T8w) forhistopathological evaluation. In addition, at the beginning (T0), end oftreatment (T8w) haemogram analysis, liver function testing and renalfunction testing were done to document the safety profile of thetreatments administered.

Histopathology of the skin biopsy was done by an expert pathologist andthe stratum granulosum parameter was measured at visits T₀ and T_(8w).The results of the stratum granulosum measurements were scored asfollows:

-   -   (3)=representing the absence of a granular layer    -   (2)=representing a thinning of the granular layer    -   (1)=representing a normal thickness granular layer of normal        thickness.        The stratum granulosum parameter represents the thickness and        integrity of the stratum granulosum layer. The more active the        disease, the thinner the stratum granulosum layer and the lower        the lipid content.

FIG. 2 presents photomicrographs of the skin of patients observed beforeand after the 8-week treatment with the herbal formulation of Wrightiatinctoria and Cocos nucifera. It is clear from the photographs that thetreatment with the herbal formulation is very effective in increasingthe integrity and thickness of the stratum granulosum layer as comparedto the condition prior to the start of treatment.

Statistical Analysis of Results

Results of the statistical analysis of the stratum granulosummeasurement data for the two treatment groups are presented in Table 3.A p-value of 0.05 is considered to be significant.

TABLE 3 Statistical Analysis of Histopathology Measurements for StratumGranulosum Herbal Allopathy Control TIME (Group I) (Group II) POINTSMEAN SD MEAN SD t P-Value T0W 2.00 0.94 1.6 0.84 1.000 0.331 T8W 1.000.00 1.2 0.63 1.000 0.331 PAIRED t 3.354 1.078 STATISTIC SIG 0.008 0.309(2 - TAILED)

To examine the treatment effects, if any, a t-test was performed withthe data taken for the two groups at the beginning and the end of thetreatment. No statistical significance was observed (p>0.05) fortreatment effects on the stratum granulosum measurements at thebeginning (p=0.388) and the end of treatment. That is, no difference invalues could be attributed to the different treatments).

To examine the time effects within each group, a paired t-test was donewith data at the beginning and end of treatment within each group. Withthe herbal group, there was a statistically significant time effect(p-values equal to 0.015) on the stratum granulosum measurements and itwas found that the stratum granulosum values decreased with timesuggesting a positive response to herbal treatment with time.

However, with the allopathy control (Group II), no statisticallysignificant time effect was found for the Allopathy control formulation(p-value equal to 0.081).

The statistical data analysis clearly indicates that the herbaltreatment for regression of stratum granulosum in keratinizationdisorders is effective and is superior to the allopathy controlformulation.

Safety Evaluation

The safety of the use of the herbal formulation over the treatmentperiod was evaluated by taking measurements of vital signs, haemogrammeasurements, liver function test (LFT) measurements, and renal functiontest (RFT) measurements and analyzing the data as a function of time.

The vital signs were measured 6 times during the treatment: T0, T1w,T2w, T4w, T6w, and T8w; the haemogram, the LFT and RFT measurements weremade only at the beginning and end of the treatment (T0, T8w).

The results of the statistical analysis of the vital sign measurements(Systolic and Diastolic BP, pulse rate and respiratory measurements) arepresented in Table 4. The BP was measured using a manual mercurysphygmomometer. The unit of measurement is mm of Hg. The pulse rate wasmeasured (beats per minute) in the radial artery by palpating the arterywith the middle, index and ring finger. The respiratory rate wasmeasured by watching the expansion of the abdomen with each respirationand counting the expansions for one minute.

TABLE 4 Statistical Analysis of Vital Sign Measurements for herbaltreatment. Respiratory Time BP-Systolic BP-Diastolic Pulse Rate RatePoints Mean SD Mean SD Mean SD Mean SD 0w 121.10 15.31 81.00 8.76 87.6017.33 23.00 6.20 T1w 111.40 11.43 77.00 8.01 75.80 8.77 21.40 7.00 T2w114.00 14.30 79.20 9.85 74.60 11.70 22.30 6.93 T4w 107.00 8.23 79.005.68 85.40 11.47 24.20 5.45 T6w 111.40 8.00 78.80 5.27 78.70 22.60 24.003.62 T8w 109.00 12.87 78.00 6.32 82.40 11.96 25.40 4.99 Grand 112.3212.36 78.83 7.28 80.75 14.88 23.38 5.72 Mean 1-Way 1.674 0.317 1.2730.612 ANOVA F-value p-value 0.157 0.901 0.289 0.691

A regular one-way ANOVA was also used to analyze the data at differenttime points for the vital signs measurements. The data clearly indicatesthat there were no statistically significant time effects on BP systolicmeasurements (p=0.157); BP diastolic measurements (p=0.901); pulse ratemeasurements (p=0.289) and respiratory rate measurements (0.691) withthe herbal treatment. In summary, there is no statistically significantchange in the vital sign measurements over time due to treatment withthe formulation of the present herbal invention for the regression ofstratum granulosum in keratinization disorders, suggesting no safetyissues.

Results of the statistical analysis of the haemogram measurements [Totalcount of white blood cells (TC), differential white blood cells count aspolymorphonuclear neutrophil (DC-P), lymphocytes (DC-L), eosinophils(DC-E) and haemoglobin (Hb)] are presented in Table 5. TC (Total countof white blood cells in the blood) was measured using a NeubauerCounting Chamber. The normal range for TC measurements is 4000-11,000cells per cubic millimeter. DC-P, which stands for the percentage ofP-polymorphonuclear neutrophil, was measured using Neubauer CountingChamber. The normal range for DC-P measurements is 55-65% of total whitecell count. DC-L, which is the percentage of lymphocytes present, wasmeasured using a Neubauer Counting Chamber. The normal range for DC-LMeasurements is 30-40% of the total white cell count. DC-L was measured.DC-E, which is the percentage of eosinophils, was measured using theNeubauer Counting Chamber. The normal range for DC-E measurements is1-7% of the total white blood cell count. DC-E was measured. HB, whichis the haemoglobin measurements, was measured using the RA 50Biochemical Analyzer and the normal range is 12-14 gms.

TABLE 5 Statistical Analysis of Haemogram Measurements for the herbaltreatment. Time TC DC-P DC-L DC-E HB Points Mean SD Mean SD Mean SD MeanSD Mean SD T0w 7343.00 1588.76 57.30 2.95 37.90 1.79 4.80 2.90 13.021.72 T8w 8634.00 1104.94 58.90 2.69 37.10 2.38 4.00 2.49 12.95 0.94Paired in −1291.00 2279.49 −1.60 3.78 0.80 3.22 0.80 3.77 0.075 2.13differ mean Paired t −1.791 −1.340 0.784 0.672 0.100 statistic Sig 0.1070.213 0.453 0.519 0.924 (2-tailed)

To examine the time effects, a paired t-test was done with data taken atthe beginning and end of treatment for each of the haemogrammeasurements. The data clearly indicates that there were nostatistically significant time effects on TC measurements (p=0.107);DC-P measurements (p=0.213); DC-L measurements (p=0.453); DC-Emeasurements (p=0.519) and HB measurements (p=0.924) with the herbaltreatment. In summary, there is no statistically significant change inhaemogram measurements with time due to the treatment with the herbalformulation, suggesting no safety issues.

Results of the statistical analysis of the liver function test (LFT)measurements [serum glutamic oxalo acetic transaminase (SGOT), serumglutamic pyruvic transaminase (SGPT) and serum bilirubin] are presentedin Table 6. SGOT, serum glutamic-oxalo acetic transaminase(international unit per liter), was measured at visits T₀ and T_(8w).The normal range is 0-46 IU/L. SGPT, serum glutamic pyruvic transaminase(international units/liter) was measured at visits T₀ and T_(8w). Thenormal SGPT ranges from 0 to 49 IU/L. The serum bilirubin was measuredat visits T₀ and T_(8w). The normal serum bilirubin ranges from 0.0 to1.0 mg/dl.

TABLE 6 Statistical Analysis of Liver Function Test (LFT) Measurementsfor the herbal treatment. Time SGOT SGPT Serum Bilirubin Points Mean SDMean SD Mean SD T0w 24.90 8.80 26.10 14.78 0.73 0.23 T8w 24.00 8.9426.60 11.01 0.69 0.24 Paired in 0.90 10.97 −0.50 11.24 0.035 0.31 differmean Paired t 0.259 0.141 0.352 statistic Sig 0.801 0.891 0.733(2-tailed)

To examine the time effects a paired t-test was done with data taken atthe beginning and the end of treatment for each of the LFT measurements.The data clearly indicates that there were no statistically significanttime effects on SGOT measurements (p=0.801); SGPT measurements(p=0.891); and the serum bilirubin measurements (p=0.733) with theherbal treatment. In summary, there is no statistically significantchange in LFT measurements with time due to treatment with the herbalformulation of the present invention for the regression of stratumgranulosum in keratinization disorders, suggesting no safety issues.

Results of the statistical analysis of the Renal Function Test (RFT)measurements for serum creatinine and serum urea,] are presented inTable 7. Serum creatinine was measured at visits T₀ and T_(8w). Thenormal serum creatinine value ranges from 0.8 to 1.4 mg/dl. Serum ureawas measured at visits T₀ and T_(8w). The normal serum urea value rangesfrom 10 to 50 mg/dl.

TABLE 7 Statistical Analysis of Renal Function Test (RFT) Measurementsfor the Herbal Treatment. Time Serum Creatinine Serum Urea Points MeanSD Mean SD T0w 1.06 0.22 32.40 17.50 T8w 1.08 0.18 25.49 7.75 Paired in−0.021 0.244 6.91 18.81 differ mean Paired t −0.271 1.161 statistic Sig0.792 0.275 (2-tailed)

To examine the time effects paired t-test was done with data at thebeginning and end of treatment for each of the RFT measurements. Thedata clearly indicates that there were no statistically significant timeeffects on serum creatinine measurements (p=0.792) and serum ureameasurements (p=0.275) with the herbal treatment. In summary, there isno statistically significant change in RFT measurements with time due totreatment with the herbal formulation of the present invention for theregression of stratum granulosum in keratinization disorders, suggestingno safety issues.

It is clear from the histopathological examination and statisticalanalysis of the clinical data that the herbal formulations of thecompositions of the present invention are very effective in thetreatment of stratum granulosum and is superior to the allopathycontrol. In addition, the evaluation of haemogram, LFT and RFT testresults clearly show that the herbal formula of the present invention isalso safe to use on humans when used as directed.

Other modifications and variations of the present invention will becomeapparent to those skilled in the art from an examination of the abovespecification and examples. Therefore, other variations of the presentinvention may be made which fall within the scope of the appended claimseven though such variations were not specifically discussed above.

1. An herbal composition that, when used topically in therapeuticallyeffective amounts, is effective for the regression of stratum granulosumin keratinization disorders, comprising: an extract of Wrightiatinctoria in a non-aqueous medium, an extract of Cocos nucifera andpharmaceutically or cosmetically acceptable excipients for use inointment, oil, soap and shampoo formulations.
 2. The herbal compositionaccording to claim 1, wherein the non-aqueous medium for the herbalextract is a non-volatile oil.
 3. The herbal composition according toclaim 2, wherein the non-volatile oil is a vegetable oil, selected fromthe group: coconut oil, gingely oil (sesame seed oil), sunflower oil,corn oil, and a refined vegetable oil.
 4. The herbal compositionaccording to claim 2, wherein the non-volatile oil is present in theextract in an amount of from 80% to 99% by weight of the extract.
 5. Theherbal composition according to claim 1, wherein the Wrightia tinctoriais obtained from at least one of the leaves, twigs, flowers, and fruitof the Wrightia tinctoria plant.
 6. The herbal composition according toclaim 5, wherein the herbal extract of Wrightia tinctoria is present inthe non-aqueous medium in an amount of from 1% to 20% by weight.
 7. Theherbal composition according to claim 1, wherein the Cocos nucifera isextracted from the copra of the coconut.
 8. The herbal compositionaccording to claim 7, wherein the Cocos nucifera is present in theamount of 40% to 80% by weight.
 9. An ointment formulation for theregression of stratum granulosum in keratinization disorders, that, whenused topically in therapeutically effective amounts, comprises theherbal composition according to claim 1 and pharmaceutically acceptableexcipients.
 10. An oil formulation for the regression of stratumgranulosum in keratinization disorders, that, when used topically intherapeutically effective amounts, comprises the herbal compositionaccording to claim 1 and pharmaceutically acceptable excipients.
 11. Aliquid soap formulation for the regression of stratum granulosum inkeratinization disorders, that, when used topically in therapeuticallyeffective amounts, comprises the herbal composition according to claim 1and pharmaceutically acceptable excipients.
 12. A shampoo formulationfor the regression of stratum granulosum in keratinization disorders,that, when used topically in therapeutically effective amounts,comprises the herbal composition according to claim 1 andpharmaceutically acceptable excipients.
 13. A method for the regressionof stratum granulosum in keratinization disorders, which comprisesadministering to a subject in need thereof a formulation comprising atherapeutically effective amount of a composition according to claim 1,said formulation in a form chosen from the group: an oil, a shampoo, anointment and a liquid soap.
 14. An herbal composition for the regressionof stratum granulosum in psoriatic lesions when used topically intherapeutically effective amounts, comprising: an extract of Wrightiatinctoria in a non-aqueous medium, an extract of Cocos nucifera andpharmaceutically or cosmetically acceptable excipients for use inointment, oil, soap and shampoo formulations.
 15. The herbal compositionaccording to claim 13, wherein the non-aqueous medium for the herbalextract is a non-volatile oil.
 16. The herbal composition according toclaim 14, wherein the non-volatile oil is a vegetable oil, selected fromthe group: coconut oil (Cocos nucifera), gingely oil (sesame seed oil),sunflower oil, corn oil, and a refined vegetable oil.
 17. The herbalcomposition according to claim 14, wherein the non-volatile oil ispresent in the extract in an amount of from 80% to 99% by weight of theextract.
 18. The herbal composition according to claim 14, wherein theWrightia tinctoria is obtained from at least one of the leaves, twigs,flowers, and fruit of the Wrightia tinctoria plant.
 19. The herbalcomposition according to claim 17, wherein the herbal extract ofWrightia tinctoria is present in the non-aqueous medium in an amount offrom 1% to 20% by weight.
 20. The herbal composition according to claim18, wherein the Cocos nucifera is extracted from the copra of thecoconut.
 21. The herbal composition according to claim 19, wherein theCocos nucifera is present in the amount of 40% to 80% by weight.
 22. Anointment formulation for the regression of stratum granulosum inpsoriatic lesions that, when used topically in therapeutically effectiveamounts, comprises the herbal composition according to claim 13 andpharmaceutically acceptable excipients.
 23. An oil formulation for theregression of stratum granulosum in psoriatic lesions that, when usedtopically in therapeutically effective amounts, comprises the herbalcomposition according to claim 13 and pharmaceutically acceptableexcipients.
 24. A liquid soap formulation for the regression of stratumgranulosum in psoriatic lesions that, when used topically intherapeutically effective amounts, comprises the herbal compositionaccording to claim 13 and pharmaceutically acceptable excipients.
 25. Ashampoo formulation for the regression of stratum granulosum inpsoriatic lesions that, when used topically in therapeutically effectiveamounts, comprises the herbal composition according to claim 13 andpharmaceutically acceptable excipients.
 26. A method for the regressionof stratum granulosum in psoriatic lesions which comprises administeringto a subject in need thereof a formulation comprising a therapeuticallyeffective amount of at least one composition according to claim 1, saidformulation in a form chosen from the group: an oil, a shampoo, anointment and a liquid soap.